Acute and sub-chronic toxicity bioassays of Olive Mill Wastewater on the Eastern mosquitofish Gambusia holbrooki.

Olive oil production generates large volumes of wastewaters mostly in peri-Mediterranean countries with adverse impacts on the biota of the receiving aquatic systems. Few studies have however documented its toxicity on aquatic species, with an almost total lack of relative studies on fish. We assessed the acute and sub-chronic OMW toxicity, as well as the acute and sub-chronic behavioural, morphological and biochemical effects of OMW exposure on the mosquitofish Gambusia holbrooki. LC50 values of the acute bioassays ranged from 7.31% (24 h) to 6.38% (96 h). Behavioural symptoms of toxicity included hypoactivity and a shift away from the water surface, coupled with a range of morphological alterations, such as skin damage, excessive mucus secretion, hemorrhages, fin rot and exophhalmia, with indications also of gill swelling and anemia. Biochemical assays showed that OMW toxicity resulted in induction of catalase (CAT) and inhibition of acetylcholinesterase (AChE) activities. The implications of our results at the level of environmental policy for the sustainable management of the olive mill industry, i.e. the effective restriction of untreated OMW disposal of in adjacent waterways, as well as the implementation of new technologies that reduce their impact (detoxification and/or revalorization of its residues) are discussed.

Water quality and benthic fauna biodiversity in a unique small wetland at Messinia, Greece.

The wetland of Aghios Floros is located in the Prefecture of Messinia (S. W. Peloponnese, Greece) and occupies a small area, covered permanentlywith water. Flooding of the surrounding area is defended by an artificial channel that discharge large quantity of water into Pamisos River in whose river basin the Aghios Floros station belongs. At the sampling site various physico-chemical and conventional pollution parameters as well as hydrochemical variables were measured during the wet and the dry period of 2011. The hydromorphological and multihabitat approach of RIVPACS method was applied in situ, which gives an overall image of the landscape. The site was classified as 'Good' according to the Greek River Nutrient Classification System (GR.NCS) and the benthic macroinvertebrate fauna assemblages that dominated the area pointed out a 'Good' biological status as well. The biotic and abiotic sample processing, carried out in compliance with the demands of the Water Framework Directive, in general revealed high ecological status of the station. Specifically, a rich diversity and abundance of some macroinvertebrate families was recorded and regarding the aquatic flora the area is dominated by the water lilies species of Nymphaea alba which are unique in the area of Peloponnese.

Stable isotopes reveal organic nitrogen pollution and cycling from point and non-point sources in a heavily cultivated (agricultural) Mediterranean river basin.

The Pinios River Basin (PRB) is the most intensively cultivated area in Greece, which hosts numerous industries and other anthropogenic activities. The analysis of water samples collected monthly for ∼1 ½ years in eight monitoring sites in the PRB revealed nitrate pollution of organic origin extending from upstream to downstream and occurring throughout the year, masking the signal from the application of synthetic fertilizers. Nitrate concentrations reached up to 3.6 mg/l as NO3--N, without exceeding the drinking water threshold of ∼11.0 mg/l (as NO3--N). However, the water quality status was "poor" or "bad" in ∼50 % of the samples based on a local index, which considers the potential impact of nitrate on aquatic biological communities. The δ15Ν-ΝΟ3- and δ18O-NO3- values ranged from +4.4 ‰ to +20.3 ‰ and from -0.5 ‰ to +14.4 ‰, respectively. The application of a Bayesian model showed that the proportional contribution of organic pollution from industries, animal breeding facilities and manure fertilizers exceeded 70 % in most river sites with an overall uncertainty of ∼0.3 (UI90 index). The δ18O-NO3- and its relationship with δ18O-H2O revealed N-cycling and mixing processes, which were difficult to identify apart from the uptake of nutrients by phytoplankton during the growing season and metabolic activities. The strong correlation of δ15Ν-ΝΟ3- values with a Land Use Index (LUI) and a Point Source Index (PSI) highlighted not only the role of non-point nitrate sources but also of point sources of nitrate pollution on water quality degradation, which are usually overlooked. The nitrification of organic wastes is the dominant nitrate source in most rivers in Europe. The systematic monitoring of rivers for nitrate isotopes will help improve the understanding of N-cycling and the impact of these pollutants on ecosystems and better inform policies for protection measures so to achieve good ecological status.

Sources of major elements and nutrients in the water cycle of an undisturbed river basin - Samothraki Island, Greece.

We studied the origin of elements of an undisturbed stream basin during the dry season as derived by atmospheric inputs and lithological processes. Α mass balance model was applied taking into account atmospheric (rain and vapor) inputs and their origin from marine aerosol and dust, as well as the contribution of rock mineral weathering and dissolution of soluble salts. The model results were enhanced using element enrichment factors, element ratios and water stable isotopes. Weathering and dissolution of bedrock and soil minerals contributed the main element portions, besides sodium and sulfate that chiefly derived from wet deposition. Vapor was shown to contribute water to inland waters of the basin. However, rain was the main source of elements compared to vapor, with marine aerosol being the only atmospheric chloride source, contributing also over 60 % of atmospheric sodium and magnesium. Silicate derived from mineral weathering (mainly plagioclase and amorphous silica), while soluble salt dissolution contributed the main portions of the rest of major elements. In headwater springs and streams, element concentrations were more affected by atmospheric inputs and silicate mineral weathering was more intense, contrary to lowland waters that were more affected by soluble salt dissolution. Effective self-purification processes were mirrored in low nutrient levels, despite the significant inputs from wet deposition, with rain being more important contributor than vapor for the majority of nutrient species. Relatively high nitrate concentrations in headwaters were attributed to increased mineralization and nitrification, while the downstream nitrate diminishing was due to prevailing denitrification processes. The ultimate goal of this study is to contribute in establishing stream elements' reference conditions using mass balance modeling approaches.

Priority and emerging organic microcontaminants in three Mediterranean river basins: Occurrence, spatial distribution, and identification of river basin specific pollutants.

There is a worldwide growing use of chemicals by our developed, industrialized, and technological society. More than 100,000 chemical substances are thus commonly used both by industry and households. Depending on the amount produced, physical-chemical properties, and mode of use, many of them may reach the environment and, notably, the aquatic receiving systems. This may result in undesirable and harmful side-effects on both the human and the ecosystem's health. Mediterranean rivers are largely different from Northern and Central European rivers in terms of hydrological regime, climate conditions (e.g. air temperature, solar irradiation, precipitation), and socio-economics (e.g. land use, tourism, crop types, etc.), with all these factors leading to differences in the relative importance of the environmental stressors, in the classes and levels of the pollutants found and their environmental fate. Furthermore, water scarcity might be critical in affecting water pollution because of the lowered dilution capacity of chemicals. This work provides raw chemical data from different families of microcontaminants identified in three selected Mediterranean rivers (the Sava, Evrotas, and Adige) collected during two sampling campaigns conducted in 2014 and 2015 in three different matrices, namely, water, sediments, and biota (fish). More than 200 organic micropollutants were analyzed, including relevant groups like pharmaceuticals, personal care products, perfluorinated compounds, pesticides, pyrethroid insecticides, flame retardants, and persistent organic pollutants. Data obtained were summarized with some basic statistics for all compound families and matrices analyzed. Observed occurrence and spatial patterns were interpreted both in terms of compound physical-chemical properties and local environmental pressures. Finally, their spatial distribution was examined and their ecotoxicological risk in the water phase was assessed. This allowed locating, at each basin, the most polluted sites ("hot spots") and identifying the respective river basin specific pollutants (RBSPs), prioritizing them in terms of the potential ecotoxicological risk posed to the aquatic ecosystems.

Unravelling the effects of multiple stressors on diatom and macroinvertebrate communities in European river basins using structural and functional approaches.

Rivers suffer from more severe decreases in species diversity compared to other aquatic and terrestrial ecosystems due to a variety of pressures related to human activities. Species provide different roles in the functioning of the ecosystem, and their loss may reduce the capacity of the ecosystems to respond to multiple stressors. The effects on diversity will differ based on the type, combination and severity of stressors, as well as on the characteristics of the community composition and tolerance. Multiple trait-based approaches (MTBAs) can help to unravel the effects of multiple stressors on communities, providing a mechanistic interpretation, and, thus, complementing traditional biodiversity assessments using community structure. We studied the relationships between diversity indexes and trait composition of macroinvertebrate and diatom communities, as well as environmental variables that described the hydrological and geomorphological alterations and toxic pollution (pesticides and pharmaceuticals) of three different European river basins: the Adige, the Sava, and the Evrotas. These river basins can be considered representative cases of different situations in European freshwater systems. Hydrological variables were the main drivers determining the community structure and function in the rivers, for both diatoms and macroinvertebrates. For diatom communities, pharmaceutical active compound (PhAC) toxic units were also identified as a very important driver of diversity changes, explaining up to 57% of the variance in taxonomic richness. For macroinvertebrates, river geomorphology was an important driver of structural changes, particularly affecting Plecoptera richness. In addition, PhAC and pesticide toxic units were also identified as stressors for macroinvertebrate communities. MTBA provided a detailed picture of the effects of the stressors on the communities and confirmed the importance of hydrological variables in shaping the functional attributes of the communities.

The environmental state of rivers in the Balkans--a review within the DPSIR framework.

Fifteen major Balkan rivers with over 80% of the inflows in Eastern Mediterranean were examined for their environmental state within the DPSIR framework. Physicogeographic and hydrochemical conditions differ substantially among river basins, which may be roughly classified into three main zones. Despite strong fragmentation, most of the rivers are liable to flash floods and have low summer flow. Decreasing precipitation and (mis)management caused a dramatic discharge reduction over the last decades. Wars, political instability, economical crises over the past decades, combined with administrative and structural constraints, poor environmental planning and inspection and, frequently, a lack of environmental awareness imposed significant pressures on rivers. Large wetland areas were drained in favour of widespread intensive agriculture. The treatment of municipal wastewaters is barely adequate in Greece and insufficient elsewhere, while management and treatment of mining and industrial wastewaters is overall poor. In general, lowland river sections are hydro-morphologically modified and are at the greatest pollution risk, while upstream areas mostly retain their natural conditions. Nutrient concentrations in a number of central and eastern Balkan rivers often exceed quality standards, whereas pesticides and heavy metals, partly of geochemical origin, occasionally exceed quality standards. Reservoirs retain vast masses of sediments, thus adversely affecting delta evolution, while dam operation disturbs the seasonal hydrological and hydrochemical regimes. Almost all Balkan countries face daunting water resource challenges because of urgently needed investments in water supply, sanitation, irrigation, and hydroelectricity. International treaties and designations and European Union Directives have mobilized pollution mitigation and conservation efforts. However, the application of environmental legislation has proved in a number of cases inadequate. Constraints arise from long-standing top-down planning traditions, inadequate planning of national environmental policies, poor administrative capacities, and heavy investment requirements, often combined with a lack of environmental awareness.

Stream invertebrate communities are primarily shaped by hydrological factors and ultimately fine-tuned by local habitat conditions.

The environmental factors that determine species richness and community structure in running waters have long been studied, but how these factors hierarchically and/or interactively influence benthic communities remains unclear. To address this research gap, we identified the principal abiotic factors that determine the taxonomic composition and functionality of stream macroinvertebrate communities and explored possible hierarchical and/or interactive patterns. We analyzed a large dataset from Greek rivers, and compared multiple macroinvertebrate metrics and traits between perennial and intermittent watercourses during wet and dry periods. We found that macroinvertebrates were primarily influenced by two ecological gradients: (i) aquatic vegetation-conductivity; and (ii) water temperature-canopy cover. Macroinvertebrates in perennial rivers were mainly influenced by the first gradient, whereas in intermittent rivers both gradients were important. Taxonomic richness and diversity were higher and temporally stable within years in perennial rivers, whereas in intermittent rivers, these metrics peaked during early summer, before the onset of streambed desiccation. The two environmental gradients determined the taxonomic richness and diversity in both spring and summer; however, a clear influence of hydrological factors (wetted width, water depth, flow velocity and discharge) was observed only in the intermittent samples. We conclude that the benthic invertebrate taxonomic richness and diversity in highly variable environments is primarily determined by hydrological variation and ultimately fine-tuned by local habitat factors. As climate change scenarios predict severe modification of hydrological and local habitat factors, this study concludes that in river management, hydrological restoration should be prioritized over other local habitat factors by maintaining natural hydrological variability, to ensure aquatic community richness and diversity.

Personal care products reconnaissance in EVROTAS river (Greece): Water-sediment partition and bioaccumulation in fish.

Twenty-six common ingredients of personal care products (PCPs) in water, sediment and fish from the Evrotas River (Greece) were investigated. Water sample analysis revealed the occurrence of twenty PCPs at concentrations ranging from 2.8 to 2031.0 ng l-1, the maximum corresponding to the endocrine disrupting UV filter benzophenone 3 (BP3). In sediment samples, six compounds were found to be adsorbed, the highest concentration being that of 4-methylbenzylidene camphor (4MBC, 1400.4 ng g-1 dw). Evrotas cyprinid fish (Squalius keadicus) showed a high accumulation potential for these chemicals, 100% detection frequency with maximum concentration that of benzophenone 2 (BP2, 41.9 ng g-1 dw). These data allowed estimating the distribution coefficients sediment-water (DCs-w) and the bioaccumulation factors (BAFs) of the pollutants investigated. Calculated rates revealed that benzophenone 1 (BP1), 4-hydroxybenzophenone (4HB) and ethyl-4-(dimethyl-amino)benzoate (EtPABA) have a strong tendency to adsorb onto the sediments, showing high DCs-w, i.e. 8.2E + 4 l g-1, 6.7E + 4 l g-1 and 5.7E + 3 l g-1, respectively. BFAs were only estimated for 5-methyl benzotriazole (MeBT), the compound having paired data from fish and water. The obtained values (range 2.0E + 2 l g-1-3.8E + 3 l g-1), indicated MeBT's strong bioaccumulation. Risk assessment of the investigated compounds for several aquatic organisms indicated a high ecological risk (HQ > 1) for BP3 and medium ecological risk (HQ ~ 0.5) for ODPABA.

Effects of olive mill wastewater discharge on benthic biota in Mediterranean streams.

Olive mill wastewaters (OMW) discharging in river ecosystems cause significant adverse effects on their water chemistry and biological communities. We here examined the effects of OMW loads in four streams of a Mediterranean basin characterized by changing flow. The diatom and macroinvertebrate community structures were compared between upstream (control) and downstream (impacted) sites receiving OMW discharge. We also tested if effects occurred at the organism level, i.e. the occurrence of deformities in diatom valves, and the sediment toxicity on the midge Chironomus riparius. We evaluated these effects through a two-year analysis, at various levels of chemical pollution and dilution capacity. The impacted sites had high phenol concentrations and organic carbon loads during and after olive mill (OM) operation, and were characterized by higher abundances of pollution-tolerant diatom and macroinvertebrate taxa. Diatom valve deformities occurred more frequently at the impacted sites. The development of C. riparius was affected by phenolic compounds and organic carbon concentrations in the sediments. The similarity in the diatom and macroinvertebrate assemblages between control and impacted sites decreased at lower flows. Diatoms were more sensitive in detecting deterioration in the biological status of OMW receiving waterways than macroinvertebrates. Our results indicate that the negative effects of OMW extended to the whole benthic community, at both assemblage and organism level.

Defining chemical status of a temporary Mediterranean River.

Although the majority of rivers and streams in the Mediterranean area are temporary, no particular attention is being paid for such systems in the Water Framework Directive (WFD). A typical temporal Mediterranean river, draining an intensively cultivated basin, was assessed for its chemical status. Elevated concentrations of nitrates and salts in river water as well as nutrients and heavy metals in river sediments have been attributed to agricultural land uses and practices and point sources of organic pollution. A scheme for the classification of the river's chemical status (within the ecological quality classification procedure) was applied by combining pollution parameters in groups according to related pressures. In light of the temporal hydrological regime and anthropogenic impacts, sediment chemical quality elements were considered, in addition to hydrochemical ones. Despite the extensive agricultural activities in the basin, the majority of the sites examined showed a good quality and only three of them were classified as moderate. For the classification of the chemical quality of temporary water bodies, there is a need to develop ecologically relevant salinity and sediment quality standards.

Assessing the ecological effects of water stress and pollution in a temporary river - Implications for water management.

Temporary rivers are dynamic and complex ecosystems that are widespread in arid and semi-arid regions, such as the Mediterranean. Biotic communities adapted in their intermittent nature could withstand recurrent drought events. However, anthropogenic disturbances in the form of water stress and chemical pollution challenge biota with unpredictable outcomes, especially in view of climate change. In this study we assess the response of the biotic community of a temporary river to environmental stressors, focusing on water stress and pollution. Towards this aim, several metrics of four biotic groups (diatoms, macrophytes, macroinvertebrates and fish) were applied. All biotic groups responded to a pollution gradient mainly driven by land use, distinct functional groups of all biota responded to water stress (a response related to the rheophilic nature of the species and their resistance to shear stress), while the combined effects of water stress and pollution were apparent in fish. Biotic groups presented a differential temporal response to water stress, where diatom temporal assemblage patterns were explained by water stress variables of short-time response (15days), while the responses of the other biota were associated to longer time periods. There were two time periods of fish response, a short (15days) and a long-time response (60-75days). When considering management decisions, our results indicate that, given the known response of river biota to pollution, biomonitoring of temporary rivers should also involve metrics that can be utilized as early warnings of water stress.

Biogeochemical processes controlling aquatic quality during drying and rewetting events in a Mediterranean non-perennial river reach.

Desiccation and re-flooding processes play a key role on hydrological features of non-perennial rivers. This study addresses the effects of these processes on the aquatic quality and unravels underlying biogeochemical processes of an intermittent river reach in southern Greece containing a spring-fed pool. Combined spatio-temporal sampling for physicochemical parameters, major ions and nutrients and high frequency automatic monitoring during a hydrological year (2010-2011) indicate that during the dry period, solute variation was controlled by "concentration" processes (i.e. evaporative concentration and high dissolved ion input from base flow sources). Metabolic and "concentration" processes appear intensified during desiccation and water temperature rise. Photosynthesis induced carbonate precipitation, while respiration increased with gradual desiccation, but did not cause carbonate dissolution. In certain cases, photosynthesis and respiration may have occurred simultaneously as a result of differing microhabitat metabolism within the same water body. However, during the entire desiccation cycle, autotrophic production exceeded respiration resulting in relatively high oxygen concentrations, even during the night. With increasing desiccation, a rise in nutrient assimilation occurred as well as ammonification and/or desorption of ammonium from sediments, with simultaneous loss of nitrate. During initial floods, flushing of carbonate phases was not significant. In contrast, initial flood events were characterized by the dissolution of very soluble salts, i.e. epsomite-type (MgSO4∗7H2O) and gypsum (CaSO4∗2H2O). Regarding sediment transport and nutrients, a 1000-times increase of suspended sediments was observed during re-flooding, while the nutrient quality degraded, particularly for N-species. Results of the current research may serve to better understand the links of hydrological and biogeochemical processes in non-perennial rivers and streams towards their efficient management and conservation.

Combined effects of water stress and pollution on macroinvertebrate and fish assemblages in a Mediterranean intermittent river.

Water stress is a key stressor in Mediterranean intermittent rivers exacerbating the negative effects of other stressors, such as pollutants, with multiple effects on different river biota. The current study aimed to determine the response of macroinvertebrate and fish assemblages to instream habitat and water chemistry, at the microhabitat scale and at different levels of water stress and pollution, in an intermittent Mediterranean river. Sampling was conducted at high and low summer discharge, at two consecutive years, and included four reaches that were targeted for their different levels of water stress and pollution. Overall, the macroinvertebrate fauna of Evrotas River indicated high resilience to intermittency, however, variation in community structure and composition occurred under acute water stress, due to habitat alteration and change in water physico-chemistry, i.e. water temperature increase. The combined effects of pollution and high water stress had, however, pronounced effects on species richness, abundance and community structure in the pollution impacted reach, where pollution sensitive taxa were almost extirpated. Fish response to drought, in reaches free of pollution, consisted of an increase in the abundance of the two small limnophilic species, coupled with their shift to faster flowing riffle habitats, and a reduction in the abundance of the larger, rheophilic species. In the pollution impacted reach, however, the combination of pollution and high water stress led to hypoxic conditions assumed to be the leading cause of the almost complete elimination of the fish assemblage. In contrast, the perennial Evrotas reaches with relatively stable physicochemical conditions, though affected hydrologically by drought, appear to function as refugia for fish during high water stress. When comparing the response of the two biotic groups to combined acute water stress and pollution, it is evident that macroinvertebrates were negatively impacted, but fish were virtually eliminated under the two combined stressors.

Managing the effects of multiple stressors on aquatic ecosystems under water scarcity. The GLOBAQUA project.

Water scarcity is a serious environmental problem in many European regions, and will likely increase in the near future as a consequence of increased abstraction and climate change. Water scarcity exacerbates the effects of multiple stressors, and thus results in decreased water quality. It impacts river ecosystems, threatens the services they provide, and it will force managers and policy-makers to change their current practices. The EU-FP7 project GLOBAQUA aims at identifying the prevalence, interaction and linkages between stressors, and to assess their effects on the chemical and ecological status of freshwater ecosystems in order to improve water management practice and policies. GLOBAQUA assembles a multidisciplinary team of 21 European plus 2 non-European scientific institutions, as well as water authorities and river basin managers. The project includes experts in hydrology, chemistry, biology, geomorphology, modelling, socio-economics, governance science, knowledge brokerage, and policy advocacy. GLOBAQUA studies six river basins (Ebro, Adige, Sava, Evrotas, Anglian and Souss Massa) affected by water scarcity, and aims to answer the following questions: how does water scarcity interact with other existing stressors in the study river basins? How will these interactions change according to the different scenarios of future global change? Which will be the foreseeable consequences for river ecosystems? How will these in turn affect the services the ecosystems provide? How should management and policies be adapted to minimise the ecological, economic and societal consequences? These questions will be approached by combining data-mining, field- and laboratory-based research, and modelling. Here, we outline the general structure of the project and the activities to be conducted within the fourteen work-packages of GLOBAQUA.

In-stream biogeochemical processes of a temporary river.

A reach at the estuary of Krathis River in Greece was used to assess how in-stream processes alter its hydrologic and biogeochemical regime. Krathis River exhibited high annual flow variability and its transmission losses become significant, especially during the dry months. These transmission losses are enhanced in chemistry due to release of nutrients from river sediments. These fluxes are significant because they correspond to 11% of the dissolved inorganic nitrogen flux of the river. Release of nitrogen species was influenced by temperature, while release of phosphate was not because phosphate levels were below the equilibrium concentration. There is a significant amount of sediments with fine composition that create "hot spot" areas in the river reach. These sediments are mobilized during the first flush events in the fall carrying with them a significant load of nutrient and suspended matter to the coastal zone. The nutrient organic content of sediments was also significant and it was studied in terms of its mineralization capacity. The capacity for mineralization was influenced by soil moisture, exhibiting significant capacity even at moisture levels of 40%. Temporary rivers are sensitive ecosystems, vulnerable to climate changes. In-stream processes play a significant role in altering the hydrology and biogeochemistry of the water and its impacts to the coastal zone.